Predicate Devices

Reference Devices

Not Found

AI/ML (Artificial Intelligence / Machine Learning)

No
The document describes a multiplexed nucleic acid test and its associated instrument and software for interpreting PCR melt curve data. There is no mention of AI or ML in the intended use, device description, or performance studies. The software interprets data based on predefined melt curve analysis, not through learning algorithms.

Therapeutic

No.
This device is a diagnostic tool, providing simultaneous qualitative detection and identification of respiratory viral and bacterial nucleic acids. Its output is intended to aid in the diagnosis of respiratory infections, but specifically states, "The results of this test should not be used as the sole basis for diagnosis, treatment, or other management decisions." It does not directly provide therapy or treatment.

Diagnostic

Yes.

The device is intended for the "simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids" which "aids in the diagnosis of respiratory infection." This directly indicates its role as a diagnostic device.

SaMD (Software as a Medical Device)

No

The device description clearly outlines a system that includes hardware components such as the FilmArray instrument with inflatable bladders, seal points, pneumatic pistons, Peltier devices, and a digital camera, in addition to the software.

IVD (In Vitro Diagnostic)

Yes, this device is an IVD (In Vitro Diagnostic).

Here's why:

Intended Use: The "Intended Use / Indications for Use" section explicitly states that the device is a "multiplexed nucleic acid test intended for use with FilmArray systems for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infections." This clearly describes a test performed in vitro (outside the body) on a biological specimen (nasopharyngeal swab) to provide diagnostic information about a patient's condition (respiratory tract infection).
Specimen Type: The device uses nasopharyngeal swabs (NPS), which are biological specimens collected from the human body.
Purpose: The purpose is to detect and identify specific viral and bacterial nucleic acids to aid in the diagnosis of respiratory infection.
Device Description: The description details a laboratory-based system that processes the specimen and performs molecular analysis (nucleic acid purification, reverse transcription, and PCR) to generate results.

All of these characteristics align with the definition of an In Vitro Diagnostic device.

PCCP (Predetermined Change Control Plan) Authorized

N/A

Overview

Intended Use / Indications for Use

The FilmArray Respiratory Panel (RP) is a multiplexed nucleic acid test intended for use with FilmArray systems for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infections. The following organism types and subtypes are identified using the FilmArray RP: Adenovirus, Coronavirus 229E, Coronavirus HKU1, Coronavirus NL63, Coronavirus OC43, Human Metapneumovirus, Influenza A subtype H1, Influenza A subtype H3, Influenza A subtype 2009 H1, Influenza B. Parainfluenza Virus 1, Parainfluenza Virus 2, Parainfluenza Virus 3, Parainfluenza Virus 4, Human Rhinovirus/Enterovirus, Respiratory Syncytial Virus, Bordetella pertussis, Chlamydophila pneumoniae, and Mycoplasma pneumoniae. The detection and identification of specific viral and bacterial nucleic acids from individuals exhibiting signs and symptoms of a respiratory infection aids in the diagnosis of respiratory infection if used in conjunction with other clinical and epidemiological information. The results of this test should not be used as the sole basis for diagnosis, treatment, or other management decisions. Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test or, lower respiratory tract infection that is not detected by a nasopharyngeal swab specimen. Positive results do not rule out co-infection with other organisms: the agent(s) detected by the Film Array RP may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible respiratory tract infection.
Due to the small number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for Bordetella pertussis, Coronavirus 229E, Coronavirus OC43, Influenza A H1, Influenza A H3, Influenza A H1-2009. Influenza B. Mycoplasma pneumoniae, Parainfluenza Virus 1, Parainfluenza Virus 2, and Parainfluenza Virus 4 were established primarily with retrospective clinical specimens. Performance characteristics for Chlamydophila pneumoniae were established primarily using contrived clinical specimens.
Due to the genetic similarity between Human Rhinovirus and Enterovirus, the FilmArray RP cannot reliably differentiate them. A positive FilmArray RP Rhinovirus/Enterovirus result should be followed-up using an alternate method (e.g., cell culture or sequence analysis).
The FilmArray RP assay for Coronavirus OC43 may cross-react with some isolates of Coronavirus HKU1. A dual positive result may be due to cross-reactivity or may indicate a coinfection.
Performance characteristics for Influenza A were established when Influenza A 2009 H1N1, A H1, and A H3 were the predominant Influenza A viruses in circulation. Performance of detecting Influenza A may vary if other Influenza A strains are circulating or a novel Influenza A virus emerges. If infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent Influenza viruses and sent to state or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

Product codes

OCC, OEM, OOU, OEP, OTG, OOI, OZX, OZY, OQW, OZZ

Device Description

The FilmArray Respiratory Panel is a multiplex nucleic acid test designed to be used with FilmArray systems. The FilmArray RP pouch contains freeze-dried reagents to perform nucleic acid purification, reverse transcription, and nested, multiplex PCR with DNA melt analysis. FilmArray RP simultaneously conducts 20 tests for the identification of respiratory pathogens from nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infections (Table 1). Results from the FilmArray RP test are available within about one hour.
A test is initiated by loading Hydration Solution and an unprocessed patient nasopharyngeal swab (NPS) specimen (i.e. specimen mixed with Sample Buffer) into the FilmArray RP pouch. The pouch contains all of the reagents required for specimen testing and analysis in a freezedried format; the addition of Hydration Solution and specimen/Sample Buffer Mix rehydrates the reagents. After the pouch is prepared, the FilmArray software guides the user though the steps of placing the pouch into the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run.
The FilmArray instrument contains a coordinated system of inflatable bladders and seal points. which act on the pouch to control the movement of liquid between the pouch blisters. When a bladder is inflated over a reagent blister, it forces liquid from the blister into connecting channels. Alternatively, when a seal is placed over a connecting channel it acts as a valve to open or close a channel. In addition, electronically controlled pneumatic pistons are positioned over multiple plungers in order to deliver the rehydrated reagents into the blisters at the appropriate times. Two Peltier devices control heating and cooling of the pouch to drive the PCR reactions and the melt curve analysis.
Nucleic acid extraction occurs within the FilmArray pouch using mechanical and chemical Ivsis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, a nested multiplex PCR is executed in two stages. During the first stage, a single, large volume, highly multiplexed reverse transcription PCR (rt-PCR) reaction is performed. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire Defense, LLC). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to the PCR products generated during the first stage PCR reaction. The 2nd stage PCR, or nested PCR, is performed in singleplex fashion in each well of the array. At the conclusion of the 2nd stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the array captures fluorescent images of the PCR reactions and software interprets the data.
The FilmArray software automatically interprets the results of each DNA melt curve analysis and combines the data with the results of the internal pouch controls to provide a test result for each organism on the panel.

Mentions image processing

A digital camera placed in front of the array captures fluorescent images of the PCR reactions and software interprets the data.

Mentions AI, DNN, or ML

Not Found

Input Imaging Modality

Not Found

Anatomical Site

nasopharyngeal swabs (NPS)

Indicated Patient Age Range

Not Found

Intended User / Care Setting

Not Found

Description of the training set, sample size, data source, and annotation protocol

Not Found

Description of the test set, sample size, data source, and annotation protocol

Specimens previously obtained during the FilmArray RP prospective clinical evaluations comprised the base of the specimen set used for testing. This set was supplemented with other archived specimens collected from external medical facilities and reference laboratories to increase the number of specimens being tested for low prevalence analytes. A total of 102 specimens were selected such that each analyte was represented 3-5 times. Contrived nasopharyngeal swab samples were used for reproducibility studies, each spiked with various concentrations of four different RP analytes.

Summary of Performance Studies (study type, sample size, AUC, MRMC, standalone performance, key results)

Clinical Performance Study:

Study Type: Clinical evaluation and comparison study.
Sample Size: 102 specimens.
Key Results:
- 100% concordance observed for 13/20 analytes across all comparisons.
- Overall Positive Percent Agreement (PPA) for all three comparisons was 96.8% or greater (100% for 16/20 analytes), with the lower bound of the two-sided 95% confidence interval (95% CI) at 92.0% or greater.
- Overall Negative Percent Agreement (NPA) for all three comparisons was 99.2% or greater (100% for 16/20 analytes), with the lower bound of the two-sided 95% CI at 99.5% or greater.
- Occasional discrepant results attributed to analyte levels below the limit of detection (LoD) or known cross-reactivity.

Low Analyte Performance Study:

Study Type: Analytical study comparing performance at low analyte levels and at LoD.
Sample Size: A titration series was performed. For LoD testing, 20 replicates for each analyte were tested on each system configuration.
Key Results:
- Amplification and detection of each analyte comparable between configurations at all concentrations in titration series testing.
- Equivalent detection on all three configurations at LoD (>95% detection for current and modified configurations and/or overlapping 2-sided 95% confidence intervals).
- Mean Tm values for all FilmArray RP assays on modified configurations were +/- 0.5°C or less compared to the same samples tested on the current configuration.

Reproducibility Study:

Study Type: Multicenter reproducibility study.
Sample Size: 90 data points per sample, per loading procedure (across three sites over five days).
Key Results:
- The test results for FilmArray RP on FilmArray 2.0 following syringe and injection vial loading procedures were highly reproducible.
- Results were consistent with data collected on the current, single-instrument FilmArray.
- Tm reproducibility was also evaluated by site/system and overall, showing consistent mean Tm values with low standard deviations.

Key Metrics (Sensitivity, Specificity, PPV, NPV, etc.)

Clinical Performance:

Overall Positive Percent Agreement (PPA): 96.8% or greater (100% for 16/20 analytes), with 95% CI lower bound at 92.0% or greater.
Overall Negative Percent Agreement (NPA): 99.2% or greater (100% for 16/20 analytes), with 95% CI lower bound at 99.5% or greater.

Low Analyte (LoD) Testing (Detection Rate):

Adenovirus: 100%
Bordetella pertussis: 100%
Chlamydophila pneumoniae: Current System (Syringe) 95%, Modified System (Syringe) 85%, Modified System (Injection Vial) 100%
Coronavirus 229E: 100%
Coronavirus HKU1: 100%
Coronavirus NL63: 100%
Coronavirus OC43: Current System (Syringe) 95%, Modified System (Syringe) 90%, Modified System (Injection Vial) 100%
Human Metapneumovirus: Current System (Syringe) 90%, Modified System (Syringe) 75%, Modified System (Injection Vial) 95%
Human Rhinovirus: 100%
Enterovirus: 100%
Influenza A H1: 100%
Influenza A H1-2009: 100%
Influenza A H3: 100%
Influenza B: Current System (Syringe) 90%, Modified System (Syringe) 90%, Modified System (Injection Vial) 100%
Mycoplasma pneumoniae: 100%
Parainfluenza Virus 1: 100%
Parainfluenza Virus 2: 100%
Parainfluenza Virus 3: Current System (Syringe) 100%, Modified System (Syringe) 95%, Modified System (Injection Vial) 100%
Parainfluenza Virus 4: Current System (Syringe) 100%, Modified System (Syringe) 100%, Modified System (Injection Vial) 95%
Respiratory Syncytial Virus: 100%

Reproducibility (Overall % Agreement with Expected Results):

Multi-instrument + Syringe (All Sites/Systems):
- Bordetella pertussis (Moderate Positive): 100% (95.9-100%)
- Bordetella pertussis (Low Positive): 95.6% (89.0-98.8%)
- Bordetella pertussis (Negative): 100% (96.0-100%)
- Adenovirus (Moderate Positive): 97.8% (92.2%-99.7%)
- Adenovirus (Low Positive): 96.6% (90.5%-99.3%)
- Adenovirus (Negative): 100% (96.0-100%)
- Influenza A H1N1-2009 (Moderate Positive): 100% (95.9-100%)
- Influenza A H1N1-2009 (Low Positive): 100% (96.0-100%)
- Influenza A H1N1-2009 (Negative): 100% (96.0-100%)
- Respiratory Syncytial Virus Type A (Moderate Positive): 100% (96.0-100%)
- Respiratory Syncytial Virus Type A (Low Positive): 100% (95.9-100%)
- Respiratory Syncytial Virus Type A (Negative): 100% (96.0-100%)
Multi-instrument + Injection Vial (All Sites/Systems):
- Bordetella pertussis (Moderate Positive): 98.9% (94.0-100%)
- Bordetella pertussis (Low Positive): 100% (96.0-100%)
- Bordetella pertussis (Negative): 100% (96.0-100%)
- Adenovirus (Moderate Positive): 98.9% (94.0-100%)
- Adenovirus (Low Positive): 98.9% (94.0-100%)
- Adenovirus (Negative): 100% (96.0-100%)
- Influenza A H1N1-2009 (Moderate Positive): 100% (96.0-100%)
- Influenza A H1N1-2009 (Low Positive): 100% (96.0-100%)
- Influenza A H1N1-2009 (Negative): 100% (96.0-100%)
- Respiratory Syncytial Virus Type A (Moderate Positive): 100% (96.0-100%)
- Respiratory Syncytial Virus Type A (Low Positive): 100% (96.0-100%)
- Respiratory Syncytial Virus Type A (Negative): 100% (96.0-100%)
Single-instrument + Syringe (All Sites):
- Bordetella pertussis (Moderate Positive): 100% (94.0-100%)
- Bordetella pertussis (Low Positive): 100% (94.0-100%)
- Bordetella pertussis (Negative): 100% (99.3-100%)
- Adenovirus (Moderate Positive): 100% (94.0-100%)
- Adenovirus (Low Positive): 100% (94.0-100%)
- Adenovirus (Negative): 100% (99.3-100%)
- Influenza A H1N1-2009 (Moderate Positive): 100% (94.0-100%)
- Influenza A H1N1-2009 (Low Positive): 100% (94.0-100%)
- Influenza A H1N1-2009 (Negative): 100% (99.3-100%)
- Respiratory Syncytial Virus Type A (Moderate Positive): 100% (98.0-100%)
- Respiratory Syncytial Virus Type A (Low Positive): 100% (97.0-100%)
- Respiratory Syncytial Virus Type A (Negative): 100% (99.0-100%)

Predicate Device(s)

K123620

Reference Device(s)

Not Found

Predetermined Change Control Plan (PCCP) - All Relevant Information

Not Found

Regulation Number and Section

§ 866.3980 Respiratory viral panel multiplex nucleic acid assay.

(a)
Identification. A respiratory viral panel multiplex nucleic acid assay is a qualitative in vitro diagnostic device intended to simultaneously detect and identify multiple viral nucleic acids extracted from human respiratory specimens or viral culture. The detection and identification of a specific viral nucleic acid from individuals exhibiting signs and symptoms of respiratory infection aids in the diagnosis of respiratory viral infection when used in conjunction with other clinical and laboratory findings. The device is intended for detection and identification of a combination of the following viruses:(1) Influenza A and Influenza B;
(2) Influenza A subtype H1 and Influenza A subtype H3;
(3) Respiratory Syncytial Virus subtype A and Respiratory Syncytial Virus subtype B;
(4) Parainfluenza 1, Parainfluenza 2, and Parainfluenza 3 virus;
(5) Human Metapneumovirus;
(6) Rhinovirus; and
(7) Adenovirus.
(b)
Classification. Class II (special controls). The special controls are:(1) FDA's guidance document entitled “Class II Special Controls Guidance Document: Respiratory Viral Panel Multiplex Nucleic Acid Assay;”
(2) For a device that detects and identifies Human Metapneumovirus, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Human Metapneumovirus (hMPV) Using Nucleic Acid Assays;” and
(3) For a device that detects and differentiates Influenza A subtype H1 and subtype H3, FDA's guidance document entitled “Class II Special Controls Guidance Document: Testing for Detection and Differentiation of Influenza A Virus Subtypes Using Multiplex Nucleic Acid Assays.” See § 866.1(e) for the availability of these guidance documents.

Summary

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Image /page/0/Picture/1 description: The image shows the logo for the Department of Health & Human Services - USA. The logo is a circular seal with the words "DEPARTMENT OF HEALTH & HUMAN SERVICES - USA" arranged around the perimeter. Inside the circle is an emblem featuring three stylized human profiles facing to the right, with flowing lines suggesting movement or connection.

Food and Drug Administration 10903 New Hampshire Avenue Document Control Center - WO66-G609 Silver Spring, MD 20993-0002

BIOFIRE DIAGNOSTICS, LLC KRISTEN KANACK VICE PRESIDENT OF REGULATED PRODUCTS 390 WAKARA WAY SALT LAKE CITY UT 84108

February 17, 2015

Re: K143080 Trade/Device Name: FilmArrav Respiratory Panel (RP) For Use with Multi-instrument FilmArray System Regulation Number: 21 CFR 866.3980 Regulation Name: Respiratory Viral Panel Multiplex Nucleic Acid Assay Regulatory Class: II Product Code: OCC, OEM, OOU, OEP, OTG, OOI, OZX, OZY, OQW, OZZ Dated: January 15, 2015 Received: January 16, 2015

Dear Dr. Kanack:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Parts 801 and 809); medical device reporting (reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (OS) regulation (21 CFR Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

1

If you desire specific advice for your device on our labeling regulations (21 CFR Parts 801 and 809), please contact the Division of Industry and Consumer Education at its toll-free number (800) 638 2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/Resourcesfor You/Industry/default.htm. Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to

http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

You may obtain other general information on your responsibilities under the Act from the Division of Industry and Consumer Education at its toll-free number (800) 638-2041 or (301) 796-7100 or at its Internet address

http://www.fda.gov/MedicalDevices/ResourcesforYou/Industry/default.htm.

Sincerely yours,

Uwe Scherf - S for

Sally A. Hojvat, M.Sc., Ph.D. Director Division of Microbiology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K143080

Device Name FilmArray Respiratory Panel (RP)

Indications for Use (Describe)

The FilmArray Respiratory Panel (RP) is a multiplexed nucleic acid test intended for use with FilmArray systems for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infections. The following organism types and subtypes are identified using the FilmArray RP: Adenovirus 229E, Coronavirus HKU1, Coronavirus NL63, Coronavirus OC43, Human Metapneumovirus, Influenza A subtype H1, Influenza A subtype H3, Influenza A subtype 2009 H1, Influenza Virus 1, Parainfluenza Virus 2, Parainfluenza Virus 3, Parainfluenza Virus 4, Human Rhinovirus/Enterovirus, Respiratory Syncytial Virus, Bordetella pertussis, Chlamydophila pneumoniae, and Mycoplasma pneumoniae. The detection of specific viral and bacterial nucleic acids from individuals exhibiting signs and symptoms of a respiratory infection aids in the diagnosis of respiratory infection if used in conjunction with other clinical and epidemiological information. The results of this test should not be used as the sole basis for diagnosis, treatment, or other management decisions. Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test or, lower respiratory tract infection that is not detected by a nasopharyngeal swab specimen. Positive results do not rule other organisms: the agent(s) detected by the Film Array RP may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culturescence, and radiography) may be necessary when evaluating a patient with possible respiratory tract infection.

Due to the small number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for Bordetella pertussis, Coronavirus OC43, Influenza A H1, Influenza A H3, Influenza A H1-2009, Influenza B, Mycoplasma pneumoniae, Parainfluenza Virus 1, Parainfluenza Virus 2, and Parainfluenza Virus 4 were established primarily with retrospective clinical specimens. Performance characteristics for Chlamydophila pneumoniae were established primarily using contrived clinical specimens.

Due to the genetic similarity between Human Rhinovirus, the FilmArray RP cannot reliably differentiate them. A positive FilmArray RP Rhinovirus result should be followed-up using an alternate method (e.g., cell culture or sequence analysis).

The FilmArray RP assay for Coronavirus OC43 may cross-react with some isolates of Coronavirus HKU1. A dual positive result may be due to cross-reactivity or may indicate a co-infection.

Performance characteristics for Influenza A were established when Influenza A 2009 HIN1, A H1, and A H3 were the predominant Influenza A viruses in circulation. Performance of detecting Influenza A may vary if other Influenza A strains are circulating or a novel Influenza A virus emerges. If influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent Influenza viruses and sent to state or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

Type of Use (Select one or both, as applicable)

X Prescription Use (Part 21 CFR 801 Subpart D)

Over-The-Counter Use (21 CFR 801 Subpart C)

CONTINUE ON A SEPARATE PAGE IF NEEDED.

3

This section applies only to requirements of the Paperwork Reduction Act of 1995.

DO NOT SEND YOUR COMPLETED FORM TO THE PRA STAFF EMAIL ADDRESS BELOW.

The burden time for this collection of information is estimated to average 79 hours per response, including the time to review instructions, search existing data sources, gather and maintain the data needed and complete and review the collection of information. Send comments regarding this burden estimate or any other aspect of this information collection, including suggestions for reducing this burden, to:

Department of Health and Human Services Food and Drug Administration Office of Chief Information Officer Paperwork Reduction Act (PRA) Staff PRAStaff@fda.hhs.gov

"An agency may not conduct or sponsor, and a person is not required to respond to, a collection of information unless it displays a currently valid OMB number."

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510(k) Summary BioFire Diagnostics, LLC

FilmArray Respiratory Panel (RP) for use with the FilmArray 2.0 and FilmArrav Injection Vials

Introduction: According to the requirements of 21 CFR 807.92, the following information provides sufficient detail to understand the basis for a determination of substantial equivalence.

Submitted by:

BioFire Diagnostics, LLC 390 Wakara Way Salt Lake City, UT 84108 USA

Telephone: 801-736-6354 Facsimile: 801-588-0507

Contact: Kristen Kanack, ext. 330

Date Submitted: October 24, 2014

Device Name and Classification:

Trade Name: FilmArray Respiratory Panel (RP)

Regulation Number: 21 CFR 866.3980

Classification Name: Respiratory Viral Panel Multiplex Nucleic Acid Assay

Predicate Device:

K123620 - FilmArray Respiratory Panel

Intended Use:

The FilmArray Respiratory Panel (RP) is a multiplexed nucleic acid test intended for use with FilmArray systems for the simultaneous qualitative detection and identification of multiple respiratory viral and bacterial nucleic acids in nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infections. The following organism types and subtypes are identified using the FilmArray RP: Adenovirus, Coronavirus 229E, Coronavirus HKU1, Coronavirus NL63, Coronavirus OC43, Human Metapneumovirus, Influenza A subtype H1, Influenza A subtype H3, Influenza A subtype 2009 H1, Influenza B. Parainfluenza Virus 1, Parainfluenza Virus 2, Parainfluenza Virus 3, Parainfluenza Virus 4, Human Rhinovirus/Enterovirus, Respiratory Syncytial Virus, Bordetella pertussis, Chlamydophila pneumoniae, and Mycoplasma pneumoniae. The detection and identification of specific viral and

5

bacterial nucleic acids from individuals exhibiting signs and symptoms of a respiratory infection aids in the diagnosis of respiratory infection if used in conjunction with other clinical and epidemiological information. The results of this test should not be used as the sole basis for diagnosis, treatment, or other management decisions. Negative results in the setting of a respiratory illness may be due to infection with pathogens that are not detected by this test or, lower respiratory tract infection that is not detected by a nasopharyngeal swab specimen. Positive results do not rule out co-infection with other organisms: the agent(s) detected by the Film Array RP may not be the definite cause of disease. Additional laboratory testing (e.g. bacterial and viral culture, immunofluorescence, and radiography) may be necessary when evaluating a patient with possible respiratory tract infection.

Due to the small number of positive specimens collected for certain organisms during the prospective clinical study, performance characteristics for Bordetella pertussis, Coronavirus 229E, Coronavirus OC43, Influenza A H1, Influenza A H3, Influenza A H1-2009. Influenza B. Mycoplasma pneumoniae, Parainfluenza Virus 1, Parainfluenza Virus 2, and Parainfluenza Virus 4 were established primarily with retrospective clinical specimens. Performance characteristics for Chlamydophila pneumoniae were established primarily using contrived clinical specimens.

Due to the genetic similarity between Human Rhinovirus and Enterovirus, the FilmArray RP cannot reliably differentiate them. A positive FilmArray RP Rhinovirus/Enterovirus result should be followed-up using an alternate method (e.g., cell culture or sequence analysis).

The FilmArray RP assay for Coronavirus OC43 may cross-react with some isolates of Coronavirus HKU1. A dual positive result may be due to cross-reactivity or may indicate a coinfection.

Performance characteristics for Influenza A were established when Influenza A 2009 H1N1, A H1, and A H3 were the predominant Influenza A viruses in circulation. Performance of detecting Influenza A may vary if other Influenza A strains are circulating or a novel Influenza A virus emerges. If infection with a novel Influenza A virus is suspected based on current clinical and epidemiological screening criteria recommended by public health authorities, specimens should be collected with appropriate infection control precautions for novel virulent Influenza viruses and sent to state or local health departments for testing. Viral culture should not be attempted in these cases unless a BSL 3+ facility is available to receive and culture specimens.

Device Description:

The FilmArray Respiratory Panel is a multiplex nucleic acid test designed to be used with FilmArray systems. The FilmArray RP pouch contains freeze-dried reagents to perform nucleic acid purification, reverse transcription, and nested, multiplex PCR with DNA melt analysis. FilmArray RP simultaneously conducts 20 tests for the identification of respiratory pathogens from nasopharyngeal swabs (NPS) obtained from individuals suspected of respiratory tract infections (Table 1). Results from the FilmArray RP test are available within about one hour.

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Viral Respiratory Pathogens
Adenovirus
Coronavirus 229E
Coronavirus HKU1
Coronavirus NL63
Coronavirus OC43
Human Metapneumovirus
Human Rhinovirus/Enterovirus
Influenza A
H1 subtype
H3 subtype
H1-2009 subtype
Influenza B
Parainfluenza Virus 1
Parainfluenza Virus 2
Parainfluenza Virus 3
Parainfluenza Virus 4
Respiratory Syncytial Virus
Bacterial Respiratory Pathogens
Bordetella pertussis
Chlamydophila pneumoniae
Mycoplasma pneumoniae

Table 1. Bacteria and Viruses Detected by the FilmArray Respiratory Panel

A test is initiated by loading Hydration Solution and an unprocessed patient nasopharyngeal swab (NPS) specimen (i.e. specimen mixed with Sample Buffer) into the FilmArray RP pouch. The pouch contains all of the reagents required for specimen testing and analysis in a freezedried format; the addition of Hydration Solution and specimen/Sample Buffer Mix rehydrates the reagents. After the pouch is prepared, the FilmArray software guides the user though the steps of placing the pouch into the instrument, scanning the pouch barcode, entering the sample identification, and initiating the run.

The FilmArray instrument contains a coordinated system of inflatable bladders and seal points. which act on the pouch to control the movement of liquid between the pouch blisters. When a bladder is inflated over a reagent blister, it forces liquid from the blister into connecting channels. Alternatively, when a seal is placed over a connecting channel it acts as a valve to open or close a channel. In addition, electronically controlled pneumatic pistons are positioned over multiple plungers in order to deliver the rehydrated reagents into the blisters at the appropriate times. Two Peltier devices control heating and cooling of the pouch to drive the PCR reactions and the melt curve analysis.

Nucleic acid extraction occurs within the FilmArray pouch using mechanical and chemical Ivsis followed by purification using standard magnetic bead technology. After extracting and purifying nucleic acids from the unprocessed sample, a nested multiplex PCR is executed in two stages. During the first stage, a single, large volume, highly multiplexed reverse transcription PCR (rt-PCR) reaction is performed. The products from first stage PCR are then diluted and combined with a fresh, primer-free master mix and a fluorescent double stranded DNA binding dye (LC Green® Plus, BioFire Defense, LLC). The solution is then distributed to each well of the array. Array wells contain sets of primers designed specifically to amplify sequences internal to

7

the PCR products generated during the first stage PCR reaction. The 2nd stage PCR, or nested PCR, is performed in singleplex fashion in each well of the array. At the conclusion of the 2ª0 stage PCR, the array is interrogated by melt curve analysis for the detection of signature amplicons denoting the presence of specific targets. A digital camera placed in front of the array captures fluorescent images of the PCR reactions and software interprets the data.

The FilmArray software automatically interprets the results of each DNA melt curve analysis and combines the data with the results of the internal pouch controls to provide a test result for each organism on the panel.

Substantial Equivalence:

The FilmArray Respiratory Panel for use with FilmArray 2.0 and FilmArray Injection Vials is substantially equivalent to the FilmArray Respiratory Panel (K123620), which was cleared for use with the FilmArray on February 11, 2013 and determined to be a Class II device.

The following table compares the FilmArray Respiratory Panel for use with FilmArray 2.0 and FilmArray Injection Vials to the previously cleared FilmArray Respiratory Panel (K123620). The table outlines the similarities and differences between the two devices.

| Element | Predicate:
FilmArray Respiratory Panel
(K123620) | New Device:
FilmArray Respiratory Panel for use
with FilmArray System 2.0 and
FilmArray Injection Vials |
|-----------------------------|-----------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------|
| Organisms
Detected | Influenza A, Influenza A subtype H1,
Influenza A subtype H3, Influenza A
subtype 2009 H1, Influenza B, Respiratory
Syncytial Virus, Human Metapneumovirus,
Adenovirus, Parainfluenza 1, Parainfluenza
2, Parainfluenza virus 3, Parainfluenza 4,
Human Rhinovirus/Enterovirus,
Coronavirus HKU1, Coronavirus NL63,
Coronavirus 229E, Coronavirus OC43,
Mycoplasma pneumoniae, Chlamydophila
pneumoniae, and Bordetella pertussis. | Same |
| Analyte | RNA/DNA | Same |
| Specimen Types | Nasopharyngeal swabs (NPS) | Same |
| Technological
Principles | Nested multiplex RT-PCR followed by high
resolution melting analysis to confirm
identity of amplified product. | Same |
| Instrumentation | FilmArray | FilmArray or FilmArray System 2.0 |
| Time to result | About 1 hour | Same |
| Test
Interpretation | Automated test interpretation and report
generation. User cannot access raw data. | Same |

Table 2. Comparison of the FilmArray Respiratory Panel on FilmArrav 2.0 and with FilmArrav Injection Vials to the FilmArray Respiratory Panel on FilmArray (Predicate).

8

| Element | Predicate:
FilmArray Respiratory Panel
(K123620) | New Device:
FilmArray Respiratory Panel for use
with FilmArray System 2.0 and
FilmArray Injection Vials |
|--------------------------------------------|----------------------------------------------------------------------------------------------------------------------------------|------------------------------------------------------------------------------------------------------------------|
| Reagent
Hydration and
Sample Loading | Syringe-based loading procedure | Syringe-based loading procedure or
FilmArray Injection Vial-based loading
procedure |
| Sample
Preparation
Method | Sample Processing is automated in the
FilmArray RP pouch. | Same |
| Reagent Storage | Reagents are stored at room temperature. | Same |
| Controls | Two controls are included in each reagent
pouch to control for sample processing and
both stages of PCR and melt analysis. | Same |
| User
Complexity | Moderate/Low | Same |

Summary of Performance Data

Clinical Performance

The original FilmArray Respiratory Panel was developed for use with the current, single instrument FilmArray and a syringe-based pouch loading procedure. A clinical study was conducted to compare the performance observed when testing clinical specimens using the FilmArray RP in its current configuration on the current system to results obtained when testing with the modified system using the current loading tools (platform comparison) as well as results on the modified platform when syringes or FilmArray Injection Vial loading tools are used (loading tools comparison). Data obtained with the current system/tools were also compared to the modified system with FilmArray Injection Vials (multifactor comparison).

Specimens previously obtained during the FilmArray RP prospective clinical evaluations comprised the base of the specimen set used for testing. This set was supplemented with other archived specimens collected from external medical facilities and reference laboratories to increase the number of specimens being tested for low prevalence analytes. A total of 102 specimens were selected such that each analyte was represented 3-5 times.

System performance for testing these 102 specimens on each platform was calculated. For the current system, a total of 108 runs were attempted, 104 of which were completed (96.3%; 104/108). There were two run failures each for software (1.9%) and instrument (1.9%) errors. No control failures were observed. Two specimens were retested due to Influenza A 'equivocal' results.

For the modified system (paired with syringe and FilmArray Injection Vial loading) a total of 205 runs were attempted, all of which were completed (100%; 205/205). There were no control failures. One specimen tested with the FilmArray Injection Vial loading tools was retested due to an Influenza A 'equivocal' result. All specimens were of sufficient volume that retesting was possible in order to obtain valid runs for all testing configurations.

9

As shown in Table 3, 100% concordance was observed for most analytes (13/20) across all comparisons. Occasional discrepant results were observed where an analyte was detected by one or two out of three runs; in all cases the discrepant results were attributed to analyte levels below the limit of detection (LoD) for a particular assay in specimens that had previously been characterized as positive for that analyte, or due to a known cross-reactivity (discussed further in Table 3). Overall Positive Percent Agreement (PPA) for all three comparisons was 96.8% or greater (100% for 16/20 analytes), with the lower bound of the two-sided 95% confidence interval (95% CI) at 92.0% or greater. Overall Negative Percent Agreement (NPA) for all three comparisons was 99.2% or greater (100% for 16/20 analytes), with the lower bound of the twosided 95% CI at 99.5% or greater.

10

| | MS+S vs CS+S
(System Comparison) | | | MS+F vs MS+S
(Loading Tools Comparison) | | | MS+F vs CS+S
(Multifactor Comparison) | | | | | |
|------------------------------|-------------------------------------|-------|-----------|--------------------------------------------|-----------|-------|------------------------------------------|------------|------------|-------|-----------|------------|
| Analyte | PPA | % | NPA | % | PPA | % | NPA | % | PPA | % | NPA | % |
| Adenovirus | 5/5 | 100% | 97/97 | 100% | 5/5 | 100% | 97/97 | 100% | 5/5 | 100% | 97/97 | 100% |
| Coronavirus 229E | 5/5 | 100% | 97/97 | 100% | 5/5 | 100% | 97/97 | 100% | 5/5 | 100% | 97/97 | 100% |
| Coronavirus HKU1 | 6/6 | 100% | 95/96a | 99% | 7/7 | 100% | 95/95 | 100% | 6/6 | 100% | 95/96a | 99% |
| Coronavirus NL63 | 6/6 | 100% | 96/96 | 100% | 6/6 | 100% | 96/96 | 100% | 6/6 | 100% | 96/96 | 100% |
| Coronavirus OC43 | 4/5b | 80% | 97/97 | 100% | 4/4 | 100% | 98/98 | 100% | 4/5b | 80% | 97/97 | 100% |
| Human Metapneumovirus | 5/5 | 100% | 97/97 | 100% | 5/5 | 100% | 97/97 | 100% | 5/5 | 100% | 97/97 | 100% |
| Human Rhinovirus/Enterovirus | 8/10c | 80% | 92/92 | 100% | 8/8 | 100% | 92/94c | 97.9% | 9/10c | 90% | 91/92c | 98.9% |
| Influenza A | 16/16 | 100% | 86/86 | 100% | 16/16 | 100% | 86/86 | 100% | 16/16 | 100% | 86/86 | 100% |
| Influenza A H1 | 3/3 | 100% | 99/99 | 100% | 3/3 | 100% | 99/99 | 100% | 3/3 | 100% | 99/99 | 100% |
| Influenza A H1-2009 | 6/6 | 100% | 96/96 | 100% | 6/6 | 100% | 96/96 | 100% | 6/6 | 100% | 96/96 | 100% |
| Influenza A H3 | 7/7 | 100% | 95/95 | 100% | 7/7 | 100% | 95/95 | 100% | 7/7 | 100% | 95/95 | 100% |
| Influenza B | 5/5 | 100% | 97/97 | 100% | 5/5 | 100% | 97/97 | 100% | 5/5 | 100% | 97/97 | 100% |
| Parainfluenza Virus 1 | 7/7 | 100% | 95/95 | 100% | 7/7 | 100% | 95/95 | 100% | 7/7 | 100% | 95/95 | 100% |
| Parainfluenza Virus 2 | 6/6 | 100% | 96/96 | 100% | 6/6 | 100% | 95/96d | 99% | 6/6 | 100% | 95/96d | 99% |
| Parainfluenza Virus 3 | 6/6 | 100% | 96/96 | 100% | 6/6 | 100% | 96/96 | 100% | 6/6 | 100% | 96/96 | 100% |
| Parainfluenza Virus 4 | 6/6 | 100% | 96/96 | 100% | 5/6e | 83.3% | 96/96 | 100% | 5/6e | 83.3% | 96/96 | 100% |
| Respiratory Syncytial Virus | 8/8 | 100% | 94/94 | 100% | 8/8 | 100% | 94/94 | 100% | 8/8 | 100% | 94/94 | 100% |
| Bordetella pertussis | 4/4 | 100% | 98/98 | 100% | 4/4 | 100% | 97/98f | 99% | 4/4 | 100% | 97/98f | 99% |
| Chlamydophila pneumoniae | 3/3 | 100% | 99/99 | 100% | 3/3 | 100% | 99/99 | 100% | 3/3 | 100% | 99/99 | 100% |
| Mycoplasma pneumoniae | 5/6g | 83.3% | 96/96 | 100% | 5/5 | 100% | 97/97 | 100% | 5/6g | 83.3% | 96/96 | 100% |
| Overall Agreement | 121/125 | 96.8% | 1914/1915 | 99.9% | 121/122 | 99.2% | 1914/1918 | 99.8% | 121/125 | 96.8% | 1911/1915 | 99.8% |
| 95% CI | 92.0-99.1% | | | 99.7-100% | 95.5-100% | | | 99.5-99.9% | 92.0-99.1% | | | 99.5-99.9% |

Table 3. Analyte Detections across all systems and loading tools. For all "X vs Y" headers, Y is the denominating performance less than 100% are shaded. CS + S = Current System, Syringe; MS+S = Modified System, Syringe; MS+F = Modified System, FilmArray Injection Vial

" Speciment 014111-RP-0037 was originally characterized as positive for CoV NL63 and was also unexpectedly positive for CoV HKU1 when ested on the MS+S and the MS+F but was not when tested on the CS+S.

" Specimen 014111-RP-0045 was originally characterized as positive for CV HKUI and demonstrated a known cross-reactivity with the CoV OC43 assay when tested with the CS+S only.

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S Three specimens (01411-RP-0100, and 01411-RP-0101) demonstrated differential detection of HRV/EV across all testing onligurations; HRV/EV had not been reported as present in these specimens by the source laboratory.

d Speciment 014111-RP-0091 was originally characterized as positive for COV 229E and was also unexpectedly positive for PV2 when tested on the MS+F but not when tested on the CS+S or MS+S.

° Specimen 014111-RP-0101 was originally characterized as positive for B. permasis and was also unexpectedly positive for PIV4 when tested on the CS+S and the MS+S but was not when tested on the MS+F.

' Specimen 01411-RP-0097 was originally characterized as positive for B. pertussis and this and this and on the MS+F but was not detected when tested on the CS+S or MS+S.

8 Specimen 014111-RP-0073 was originally characterized as positive for M. pneumoniae and this analyte was not detected when tested on the MS+S or MS+F

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Selected Analytical Studies

Low Analyte

A comparison of performance at low analyte levels between the current FilmArray system (one instrument to one computer configuration) and the FilmArray 2.0 system (modified; up to eight instruments to one computer) was performed for FilmArray RP. A comparison of performance between the current syringe-based pouch loading procedure and a modified, injection vial-based pouch loading procedure was also performed. The purpose was to establish that FilmArray RP performance is equivalent for all system and pouch loading combinations.

Testing consisted of a titration of samples containing RP analytes at concentrations above, at, and below (10x, 1x, 0.1× and 0.01×) LoD. Additional side-by-side testing at LoD (20 replicates on each system) was performed to further demonstrate consistency between the current (syringe) and modified (syringe or injection vial) system and pouch loading combinations.

In the titration series testing, amplification and detection of each analyte were found to be comparable between configurations at all concentrations. Testing of additional replicates at LoD (Table 4) also revealed equivalent detection on all three configurations (>95% detection for the current and modified configurations and/or overlapping 2-sided 95% confidence intervals).

Respiratory Panel Analyte	Current System (Syringe) # Detected % Positive [95% CI]	Modified System (Syringe) # Detected % Positive [95% CI]	Modified System (Injection Vial) # Detected % Positive [95% CI]
Adenovirus	20/20
100%
[83.2-100%]	20/20
100%
[83.2-100%]	20/20
100%
[83.2-100%]
Bordetella pertussis	20/20
100%
[83.2-100%]	20/20
100%
[83.2-100%]	20/20
100%
[83.2-100%]
Chlamydophila pneumoniae	19/20
95%
[75.1-99.9%]	17/20
85%a
[62.1-96.8%]	20/20
100%
[83.2-100%]
Coronavirus 229E	20/20
100%
[83.2-100%]	20/20
100%
[83.2-100%]	20/20
100%
[83.2-100%]
Coronavirus HKU1b	5-May
100%
[47.8-100%]	5-May
100%
[47.8-100%]	5-May
100%
[47.8-100%]
Coronavirus NL63	20/20
100%
[83.2-100%]	20/20
100%
[83.2-100%]	20/20
100%
[83.2-100%]
Coronavirus OC43	19/20
95%
[75.1-99.9%]	18/20
90%a
[68.3-98.8%]	20/20
100%
[83.2-100%]
Human Metapneumovirus	18/20	15/20	19/20
	Current System
(Syringe)	Modified System
(Syringe)	Modified System
(Injection Vial)
Respiratory Panel Analyte	# Detected	# Detected	# Detected
	% Positive	% Positive	% Positive
	[95% CI]	[95% CI]	[95% CI]
	90%a	75%a	95%
	[68.3-98.8%]	[50.9-91.3%]	[75.1-99.9%]
	20/20	20/20	20/20
Human Rhinovirus	100%	100%	100%
	[83.2-100%]	[83.2-100%]	[83.2-100%]
	20/20	20/20	20/20
Enterovirus	100%	100%	100%
	[83.2-100%]	[83.2-100%]	[83.2-100%]
	20/20	20/20	20/20
Influenza A H1	100%	100%	100%
	[83.2-100%]	[83.2-100%]	[83.2-100%]
	20/20	20/20	20/20
Influenza A H1-2009	100%	100%	100%
	[83.2-100%]	[83.2-100%]	[83.2-100%]
	20/20	20/20	20/20
Influenza A H3	100%	100%	100%
	[83.2-100%]	[83.2-100%]	[83.2-100%]
	18/20	18/20	20/20
Influenza B	90%a	90%a	100%
	[68.3-98.8%]	[68.3-98.8%]	[83.2-100%]
	20/20	20/20	20/20
Mycoplasma. pneumoniae	100%	100%	100%
	[83.2-100%]	[83.2-100%]	[83.2-100%]
	20/20	20/20	20/20
Parainfluenza Virus 1	100%	100%	100%
	[83.2-100%]	[83.2-100%]	[83.2-100%]
	20/20	20/20	20/20
Parainfluenza Virus 2	100%	100%	100%
	[83.2-100%]	[83.2-100%]	[83.2-100%]
	20/20	19/20	20/20
Parainfluenza Virus 3	100%	95%	100%
	[83.2-100%]	[75.1-99.9%]	[83.2-100%]
	20/20	20/20	19/20
Parainfluenza Virus 4	100%	100%	95%
	[83.2-100%]	[83.2-100%]	[75.1-99.9%]
	20/20	20/20	20/20
Respiratory Syncytial Virus	100%	100%	100%
	[83.2-100%]	[83.2-100%]	[83.2-100%]

Table 4. Results of Replicate Testing at LoD for the Respiratory Panel (RP) on Current and Modified FilmArray Systems with Svringe and/or FilmArray Injection Vial Pouch Loading Procedures

BioFire Diagnostics, LLC 510(k) Multi-instrument FilmArray Respiratory Panel

13

4 Detection rate is lower than the expected 95% at LoD, but detection is comparable between the current and modified configuration(s) and 95% confidence intervals are overlapping.

° Due to insufficient volume of Coronavirus HKU1, replicate testing was not performed. Results are from titration testing at LoD.

Tm values from the LoD replicate samples were compared to assess whether Tm data are equivalent between the current and modified FilmArray systems, the syringe and injection vial

BioFire Diagnostics, LLC 510(k) Multi-instrument FilmArray Respiratory Panel

14

pouch loading procedures, and between the current and modified system/loading combined. Normal Tm variation of the current FilmArray configuration is ±0.5℃ and it was observed that mean Tm values for all FilmArray RP assays on the modified configurations were ± 0.5℃ or less compared to the same samples tested on the current configuration (△Tm System, ΔTm Loading, and △Tm Combined in Table 5).

| Organism Tm | △Tm
System | △Tm
Loading | △Tm
Combined |
|------------------------------------| | Control Control | Current (Syringe) | | | | | Control Process
Control | Current (Syringe) | | | | | Adenovirus | | | | | Adenovirus | | | | | Bordetella pertussis | | | | | Chlamydophila
pneumoniae | Cpne | | | | | Coronavirus
229E | CoV-
229E | Current (Syringe) | | | | | Coronavirus
HKU1 | CoV-
HKU1 | Current (Syringe) | | | | | Coronavirus
NL63 | CoV-
NL63 | Current (Syringe) | | | | | Coronavirus
OC43 | CoV-
OC43 | Current (Syringe) | | | | | Human
Metapneumovirus | hMPV | | | | | Human
Rhinovirus | HRV1 | | | | | Human
Rhinovirus | HRV2 | | | | | | Organism Tm | △Tm
System | △Tm
Loading | △Tm
Combined |
| | | | | | | | | | | | | | | | | | Enterovirus | | | | | | | | | | | | | | | | Influenza A
HI | FluA-
pan1
FluA-
pan2 | Current (Syringe) | 83.9 | | | | | | | | | | | | | | | | | Influenza A | | H1-2009 | | | | | | | | | | | | Influenza A | нз | | | | | | | | | Influenza B | | | Organism | Mycoplasma pneumoniae | Mpne | | | | | Parainfluenza Virus 1 | PIV1 | | | | | Parainfluenza Virus 2 | PIV2 | | | | | Parainfluenza Virus 3 | PIV3 | | | | | Parainfluenza Virus 4 | PIV4 | | | | | Respiratory Syncytial Virus | RSV | | | | | Assay | Configuration | Mean
--------------------------------|---------------------------|------------|---------------|----------------|-----------------|
| PCR
| 75.7 | -0.1 | | |
| Modified (Syringe) | 75.8 | | 0.1 | 0.0 |
| Modified (Injection Vial) | 75.7 | | | |
| RNA
| 81.7 | 0.1 | | |
| Modified (Syringe) | 81.6 | | 0.0 | 0.1 |
| Modified (Injection Vial) | 81.6 | | | |
| Adeno | Current (Syringe) | 83.6 | 0.1 | | |
| Modified (Syringe) | 83.5 | | -0.3 | -0.2 |
| Modified (Injection Vial) | 83.8 | | | |
| Adeno2 | Current (Syringe) | 87.8 | 0.2 | | |
| Modified (Syringe) | 87.6 | | -0.5 | -0.3 |
| Modified (Injection Vial) | 88.1 | | | |
| Bper | Current (Syringe) | 88.1 | -0.1 | | |
| Modified (Syringe) | 88.2 | | 0.2 | 0.1 |
| Modified (Injection Vial) | 88.0 | | | |
| Current (Syringe) | 79.3 | 0 | | |
| Modified (Syringe) | 79.3 | | 0.1 | 0.1 |
| Modified (Injection Vial) | 79.2 | | | |
| 80.7 | 0.1 | | |
| Modified (Syringe) | 80.6 | | -0.4 | -0.3 |
| Modified (Injection Vial) | 81.0 | | | |
| 75.2 | 0.1 | | |
| Modified (Syringe) | 75.1 | | 0.1 | 0.2 |
| Modified (Injection Vial) | 75.0 | | | |
| 79.8 | -0.1 | | |
| Modified (Syringe) | 79.9 | | 0.1 | 0 |
| Modified (Injection Vial) | 79.8 | | | |
| 80.4 | -0.1 | | |
| Modified (Syringe) | 80.5 | | 0.1 | 0 |
| Modified (Injection Vial) | 80.4 | | | |
| Current (Syringe) | 77.3 | -0.2 | | |
| Modified (Syringe) | 77.5 | | 0.2 | 0 |
| Modified (Injection Vial) | 77.3 | | | |
| Current (Syringe) | 83.5 | 0.2 | | |
| Modified (Syringe) | 83.3 | | -0.1 | 0.1 |
| Modified (Injection Vial) | 83.4 | | | |
| Current (Syringe) | 83.4 | 0.2 | | |
| Modified (Syringe) | 83.2 | | -0.2 | 0 |
| Modified (Injection Vial) | 83.4 | | | |
| HRV3 | Current (Syringe) | 82.8 | 0.2 | | -0.1 |
| Assav | Configuration | Mean
| Modified (Syringe) | 82.6 | | | |
| Modified (Injection Vial) | 82.9 | | -0.3 | |
| Current (Syringe) | 83.8 | 0.3 | | |
| HRV4 | Modified (Syringe) | 83.5 | | 0 | 0.3 |
| Modified (Injection Vial) | 83.5 | | | |
| Current (Syringe) | 86.5 | -0.1 | | |
| Entero1 | Modified (Syringe) | 86.6 | | 0.2 | 0.1 |
| Modified (Injection Vial) | 86.4 | | | |
| Current (Syringe) | 86.4 | -0.2 | | |
| Entero2 | Modified (Syringe) | 86.6 | | 0.2 | 0 |
| Modified (Injection Vial) | 86.4 | | | |
| Current (Syringe) | 86.1 | 0.4 | | |
| HRV3 | Modified (Syringe) | 85.7 | | 0 | 0.4 |
| Modified (Injection Vial) | 85.7 | | | |
| Current (Syringe) | 85.2 | -0.2 | | |
| HRV4 | Modified (Syringe) | 85.4 | | 0.2 | 0 |
| Modified (Injection Vial) | 85.2 | | | |
| FluA- | Current (Syringe) | 78.0 | 0.2 | | |
| H1-pan | Modified (Syringe) | 77.8 | | -0.1 | 0.1 |
| Modified (Injection Vial) | 77.9 | | | |
| 0.1 | | |
| Modified (Syringe) | 83.8 | | -0.4 | -0.3 |
| Modified (Injection Vial) | 84.2 | | | |
| Current (Syringe) | 79.8 | 0.1 | | |
| Modified (Syringe) | 79.7 | | -0.3 | -0.2 |
| Modified (Injection Vial) | 80.0 | | | |
| FluA- | Current (Syringe) | 78.1 | 0.2 | | |
| H1-pan | Modified (Syringe) | 77.9 | | -0.1 | 0.1 |
| Modified (Injection Vial) | 78.0 | | | |
| FluA- | Current (Syringe) | 78.5 | 0.3 | | |
| H1-2009 | Modified (Syringe) | 78.2 | | -0.4 | -0.1 |
| Modified (Injection Vial) | 78.6 | | | |
| FluA- | Current (Syringe) | 84.5 | 0.2 | | |
| pan l | Modified (Syringe) | 84.3 | | -0.4 | -0.2 |
| Modified (Injection Vial) | 84.7 | | | |
| FluA- | Current (Syringe) | 80.3 | 0.3 | | |
| pan2 | Modified (Syringe) | 80.0 | | -0.4 | -0.1 |
| Modified (Injection Vial) | 80.4 | | | |
| FluA- | Current (Syringe) | 81.9 | -0.1 | | |
| нз | Modified (Syringe) | 82.0 | | 0.1 | 0 |
| Modified (Injection Vial) | 81.9 | | | |
| FluA- | Current (Syringe) | 84.6 | -0.2 | | |
| pan 1 | Modified (Syringe) | 84.8 | | 0.2 | 0 |
| Modified (Injection Vial) | 84.6 | | | |
| FluA- | Current (Syringe) | 79.2 | -0.1 | | |
| pan2 | Modified (Syringe) | 79.3 | | 0.1 | 0 |
| Modified (Injection Vial) | 79.2 | | | |
| Current (Syringe) | 79.8 | 0 | | |
| FluB | Modified (Syringe) | 79.8 | | 0 | 0 |
| Modified (Injection Vial) | 79.8 | | | |
| Assay | Configuration | Mean Tm | ΔTm System | ΔTm Loading | ΔTm Combined |
| Current (Syringe) | 77.4 | 0.3 | | |
| Modified (Syringe) | 77.1 | | -0.4 | -0.1 |
| Modified (Injection Vial) | 77.5 | | | |
| Current (Syringe) | 78.0 | 0.1 | | |
| Modified (Syringe) | 77.9 | | -0.4 | -0.3 |
| Modified (Injection Vial) | 78.3 | | | |
| Current (Syringe) | 83.3 | 0.2 | | |
| Modified (Syringe) | 83.1 | | -0.4 | -0.2 |
| Modified (Injection Vial) | 83.5 | | | |
| Current (Syringe) | 80.7 | -0.1 | | |
| Modified (Syringe) | 80.8 | | 0.1 | 0 |
| Modified (Injection Vial) | 80.7 | | | |
| Current (Syringe) | 76.9 | -0.2 | | |
| Modified (Syringe) | 77.1 | | 0.4 | 0.2 |
| Modified (Injection Vial) | 76.7 | | | |
| Current (Syringe) | 80.4 | 0 | | |
| Modified (Syringe) | 80.4 | | -0.3 | -0.3 |
| Modified (Injection Vial) | 80.7 | | | |

Table 5. Comparison of Mean Tm Values for FilmArray RP Analytes on the Current and Modified Systems with Syringe or FilmArray Injection Vial Pouch Loading Procedures

15

16

Reproducibility

A multicenter reproducibility study was performed to determine between-site/system and overall reproducibility of the FilmArray Respiratory Panel (RP) on multi-instrument FilmArray 2.0 systems using the current (syringe) and modified (injection vial) pouch loading procedures.

Reproducibility testing occurred at three test sites using a panel of contrived nasopharyngeal swab samples, each spiked with various concentrations of four different RP analytes. Each analyte was evaluated at three different concentrations (Negative, Low Positive, and Moderate Positive).

The study incorporated a range of potential variation introduced by up to eight different operators, three different pouch lots, and up to 11 different FilmArray 2.0 instruments per loading procedure on three different systems. A system consisted of three instruments connected to a single computer. Samples were stored frozen and tested on five different days at three testing sites (one system. A, B, or C per site) for 90 data points per sample, per loading procedure.

A summary of results (percent (%) agreement with the expected result) for each analyte (by site/system and overall) is provided in Table 6 alongside the overall % Agreement with Expected Results originally obtained on the single-instrument system.

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| | | | Multi-instrument

Syringe
Site/System | | | | Multi-instrument
Injection Vial
Site/System | | | | Single-
instrument
Syringe |
|--------------------------------------------------------------------------------|--------------------------------------------------------|--------------------|----------------------------------------------|---------------|----------------|----------------------------------------------------------|-----------------------------------------------------|----------------|----------------|----------------------------------------------------------|----------------------------------------------|
| Organism
Tested | Concentration
Tested | Expected
Result | A | B | C | All Sites/
Systems
(95%
Confidence
Interval) | A | B | C | All Sites/
Systems
(95%
Confidence
Interval) | All Sites
(95%
Confidence
Interval) |
| Bordetella
pertussis
Strain A639
Zeptometrix
0801459 | Moderate
Positive
3x LoD
$1.2x10^4$ CFU/mL | Detected | 29/29a
100% | 30/30
100% | 30/30
100% | 89/89a
100%
(95.9-100%) | 30/30
100% | 29/30
96.7% | 30/30
100% | 89/90
98.9%
(94.0-100%) | 60/60
100%
(94.0-100%) |
| | Low Positive
1x LoD
$4x10^3$ CFU/mL | Detected | 29/30
96.7% | 30/30
100% | 27/30
90.0% | 86/90
95.6%
(89.0-98.8%) | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 60/60
100%
(94.0-100%) |
| | Negative | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 540/540
100%
(99.3-100%) |
| Adenovirus
Species C
Serotype 1
Zeptometrix
0810050CF | Moderate
Positive
3x LoD
$3.0x10^2$ TCID50/mL | Detected | 30/30
100% | 30/30
100% | 28/30
93.3% | 88/90
97.8%
(92.2%-
99.7%) | 29/30
96.7% | 30/30
100% | 30/30
100% | 89/90
98.9%
(94.0-100%) | 60/60
100%
(94.0-100%) |
| | Low Positive
1x LoD
$1.0x10^2$ TCID50/mL | Detected | 28/29a
96.6% | 30/30
100% | 28/30
93.3% | 86/89a
96.6%
(90.5%-
99.3%) | 30/30
100% | 30/30
100% | 29/30
96.7% | 89/90
98.9%
(94.0-100%) | 60/60
100%
(94.0-100%) |
| | Negative | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 540/540
100%
(99.3-100%) |
| Influenza A
H1N1-2009
A/SwineNY/
03/2009
Zeptometrix
0810109CFN | Moderate
Positive
3x LoD
$3.0x10^2$ TCID50/mL | Detected | 29/29a
100% | 30/30
100% | 30/30
100% | 89/89a
100%
(95.9-100%) | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 60/60
100%
(94.0-100%) |
| | Low Positive
1x LoD
$1.0x10^2$ TCID50/mL | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 60/60
100%
(94.0-100%) |
| | Negative | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 540/540
100%
(99.3-100%) |
| Respiratory
Syncytial
Virus Type A
Zeptometrix
0810040ACF | Moderate
Positive
3x LoD
$6.0$ TCID50/mL | Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 180/180
100%
(98.0-100%) |
| | Low Positive
1x LoD
$2.0$ TCID50/mL | Detected | 29/29a
100% | 30/30
100% | 30/30
100% | 89/89a
100%
(95.9-100%) | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 120/120
100%
(97.0-100%) |
| | Negative | Not
Detected | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 30/30
100% | 30/30
100% | 30/30
100% | 90/90
100%
(96.0-100%) | 360/360
100%
(99.0-100%) |

Table 6. Reproducibility of the FilmArray Respiratory Panel Test Results on FilmArray 2.0 (Multiinstrument) and FilmArray (Single-instrument)

4 Due to failure, a valid result could not be obtained for one of the replicates, reducing the total number of replicates for Site/System A from 30 to 29 and for All Sites/Systems from 90 to 89.

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The test results obtained for the FilmArray Respiratory Panel on FilmArray 2.0 following the syringe and injection vial loading procedures were highly reproducible and are consistent with the data collected on the current, single-instrument FilmArray in the original FilmArray RP Reproducibility evaluation.

The reproducibility of Tm for each positive assay was also evaluated by site/system and overall (all sites/systems) and a summary is provided in Table 7.

| Organism

Assay		Test Level	Site/System	Tm Reproducibility		Syringe	Injection Vial
				Mean	StDev Tm	Mean	StDev Tm
Bacteria
Bordatella
pertusis
Zeptometrix
0810050CF	Bper	Moderate Pos
3x LoD
1.2x104 CFU/mL	A	88.3	±0.3	88.3	±0.2
		B	88.2	±0.2	88.1	±0.2
		C	87.8	±0.4	88.0	±0.2
		All
Sites/Systems	88.1	±0.4	88.1	±0.3
		Low Pos
1x LoD
4.0x103 CFU/mL	A	88.3	±0.2	88.4	±0.2
			B	88.2	±0.2	88.1	±0.2
			C	87.8	±0.4	88.0	±0.2
			All
Sites/Systems	88.1	±0.3	88.2	±0.2
Viruses
	FluA-H1-2009	Mod Pos
3x LoD
300 TCID50/mL	A	78.6	±0.2	78.6	±0.2
		B	78.5	±0.2	78.4	±0.2
		C	78.1	±0.3	78.2	±0.2
		All
Sites/Systems	78.4	±0.3	78.4	±0.3
		Low Pos
1x LoD
100 TCID50/mL	A	78.6	±0.2	78.6	±0.2
			B	78.5	±0.2	78.5	±0.2
			C	78.2	±0.3	78.3	±0.2
			All
Sites/Systems	78.4	±0.3	78.5	±0.2
Influenza A
H1-2009
Zeptometrix
0810109CFN	FluA-H1-pan	Mod Pos
3x LoD
300 TCID50/mL	A	78.1	±0.2	77.9	±0.2
		B	78.1	±0.2	77.9	±0.3
		C	77.7	±0.3	77.7	±0.3
		All
Sites/Systems	77.9	±0.3	77.8	±0.3
		Low Pos
1x LoD
100 TCID50/mL	A	78.4	±0.3	78.2	±0.4
			B	78.4	±0.4	78.1	±0.3
			C	78.0	±0.5	77.9	±0.3
			All
Sites/Systems	78.3	±0.4	78.1	±0.3
	FluA-pan1	Mod Pos
3x LoD
300 TCID50/mL	A	84.9	±0.2	84.9	±0.2
		B	84.7	±0.2	84.6	±0.2
		C	84.4	±0.3	84.5	±0.2
		All
Sites/Systems	84.7	±0.3	84.7	±0.2

					Table 7. Reproducibility of Tm for Positive FilmArray RP Assays on Multi-instrument FilmArray 2.0
Systems
Organism	Assav	Test Level	Site/System	Tm Reproducibility
				Syringe
Injection Vial
				Mean	StDev
Tm	Mean	StDev
Tm
		Low Pos
1× LoD
100 TCID50/mL	A	84.9	±0.2	84.9	+0.2
			B	84.7	±0.2	84.7	±0.2
			C	84.4	+0.3	84.6	+0.1
			All
Sites/Systems	84.7	+0.3	84.7	+0.2
	FluA-pan2	Mod Pos
3× LoD
300 TCID50/mL	A	80.4	±0.2	80.2	+0.4
			B	80.4	+0.1	80.1	±0.2
			C	80.1	+0.3	80.2	±0.2
			All
Sites/Systems	80.3	+0.3	80.2	+0.3
		Low Pos
1× LoD
100 TCID50/mL	A	80.5	±0.2	80.4	+0.3
			B	80.5	+0.2	80.3	±0.2
			C	80.2	+0.3	80.2	±0.2
			All
Sites/Systems	80.4	+0.3	80.3	+0.2
Adenovirus
Zeptometrix
0810050CF	Adeno	Mod Pos
3× LoD
300 TCID50/mL	A	83.7	+0.3	83.8	+0.3
			B	83.6	+0.3	83.4	+0.3
			C	83.4	+0.4	83.4	+0.3
			All
Sites/Systems	83.6	+0.4	83.5	+0.3
		Low Pos
1× LoD
100 TCID50/mL	A	84.0	+0.3	84.0	+0.3
			B	83.9	+0.3	83.6	+0.3
			C	83.7	+0.4	83.6	+0.3
			All
Sites/Systems	83.9	+0.3	83.8	+0.3
	Adeno2	Mod Pos
3× LoD
300 TCID50/mL	A	88.1	±0.2	88.2	+0.3
			B	87.9	+0.2	87.9	±0.2
			C	87.7	+0.4	87.8	±0.2
			All
Sites/Systems	87.9	+0.3	88.0	+0.3
		Low Pos
1× LoD
100 TCID50/mL	A	88.2	+0.3	88.2	±0.2
			B	88.1	+0.1	87.9	+0.3
			C	87.8	+0.4	87.8	±0.2
			All
Sites/Systems	88.0	+0.3	88.0	+0.3
Respiratory
Syncytial Virus
Zeptometrix
0810040ACF	RSV	Mod Pos
3× LoD
6 TCID50/mL	A	80.8	+0.3	80.6	+0.5
			B	80.7	+0.3	80.6	+0.3
			C	80.4	+0.3	80.6	±0.2
			All
Sites/Systems	80.6	+0.3	80.6	+0.4
		Low Pos
1× LoD
2 TCID50/mL	A	80.8	±0.2	80.6	+0.5
			B	80.8	+0.2	80.6	+0.3
			C	80.5	+0.3	80.6	+0.2
			All
Sites/Systems	80.7	+0.3	80.6	+0.3

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